title: A model for the interfacial kinetics of phospholipase D activity on long-chain lipids.
abstract: The membrane-active enzyme phospholipase D (PLD) catalyzes the hydrolysis of the phosphodiester bond in phospholipids and plays a critical role in cell signaling. This catalytic reaction proceeds on lipid-water interfaces and is an example of heterogeneous catalysis in biology. Recently we showed that planar lipid bilayers, a previously unexplored model membrane for these kinetic studies, can be used for monitoring interfacial catalytic reactions under well-defined experimental conditions with chemical and electrical access to both sides of the lipid membrane. Employing an assay that relies on the conductance of the pore-forming peptide gramicidin A to monitor PLD activity, the work presented here reveals the kinetics of hydrolysis of long-chain phosphatidylcholine lipids in situ. We have developed an extension of a basic kinetic model for interfacial catalysis that includes product activation and substrate depletion. This model describes the kinetic behavior very well and reveals two kinetic parameters, the specificity constant and the interfacial quality constant. This approach results in a simple and general model to account for product accumulation in interfacial enzyme kinetics.
authors.affiliation: Department of Bioengineering, Pennsylvania State University, University Park, Pennsylvania, USA. smajd@engr.psu.edu
authors.forename: Sheereen
authors.initials: S
authors.lastname: Majd
authors.forename: Erik C
authors.initials: EC
authors.lastname: Yusko
authors.forename: Jerry
authors.initials: J
authors.lastname: Yang
authors.forename: David
authors.initials: D
authors.lastname: Sept
authors.forename: Michael
authors.initials: M
authors.lastname: Mayer
mesh_terms: D006096:Gramicidin / Q000378:metabolism; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008954:Models, Biological; D010739:Phospholipase D / Q000378:metabolism*
chemical_list: D008051:Lipid Bilayers; D006096:Gramicidin; D010739:Phospholipase D